Human IL-15 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY247
DY247-05
Ancillary Products Available
Human IL-15 ELISA Standard Curve
1 Image
Product Details
Procedure
Citations (10)
FAQs
Supplemental Products
Reviews

Human IL-15 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For five or fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human IL-15. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

 

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

 

  

Data Example

Human IL-15 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: IL-15

IL-15 (Interleukin 15) is a cytokine that induces or enhances the differentiation, maintenance, or activation of multiple T cell subsets including NK, NKT, Th17, Treg, and CD8+ memory cells. It also induces dendritic cell differentiation and inflammatory activation, exhibits anti-tumor activity, and inhibits the deposition of lipid in adipocytes. Complexes of IL-15 with cell surface IL-15 R alpha interact with complexes of IL-2 R beta and the Common gamma Chain on adjacent cells. This transpresentation mechanism enables cells to respond to IL-15 even if they do not express IL-15 R alpha. Ligation of membrane-associated IL-15/IL-15 R alpha complexes also induces reverse signaling that promotes activation of the IL-15/IL-15 R alpha expressing cells. The activity of circulating IL-15 is limited by its association with soluble IL-15 R alpha.

Long Name:
Interleukin 15
Entrez Gene IDs:
3600 (Human); 16168 (Mouse); 25670 (Rat); 493682 (Feline)
Alternate Names:
IL15; IL-15; IL-15MGC9721; interleukin 15; interleukin-15

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

 

Citations for Human IL-15 DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

10 Citations: Showing 1 - 10
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  1. Rheumatoid arthritis bone marrow environment supports Th17 response
    Authors: E Kuca-Warna, W Kurowska, M Prochorec-, A Radzikowsk, T Burakowski, U Skalska, M Massalska, M Pleba?czyk, B Ma?dyk-Now, I S?owi?ska, R Gasik, W Ma?li?ski
    Arthritis Res. Ther., 2017;19(1):274.
    Species: Human
    Sample Types: Plasma
  2. IL15RA drives antagonistic mechanisms of cancer development and immune control in lymphocyte-enriched triple-negative breast cancers.
    Authors: Marra P, Mathew S, Grigoriadis A, Wu Y, Kyle-Cezar F, Watkins J, Rashid M, De Rinaldis E, Hessey S, Gazinska P, Hayday A, Tutt A
    Cancer Res, 2014;74(17):4908-21.
  3. Interleukin-15 contributes to the regulation of murine adipose tissue and human adipocytes.
    Authors: Barra NG, Reid S, MacKenzie R
    Obesity (Silver Spring), 2010;18(8):1601-7.
    Species: Human
    Sample Types: Complex Sample Type
  4. Human bronchial intraepithelial T cells produce interferon-gamma and stimulate epithelial cells.
    Authors: Hirosako S, Goto E, Fujii K, Tsumori K, Hirata N, Tsumura S, Kamohara H, Kohrogi H
    Clin. Exp. Immunol., 2009;155(2):266-74.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Islet transplantation in patients with autoimmune diabetes induces homeostatic cytokines that expand autoreactive memory T cells.
    Authors: Monti P, Scirpoli M, Maffi P, Ghidoli N, De Taddeo F, Bertuzzi F, Piemonti L, Falcone M, Secchi A, Bonifacio E
    J. Clin. Invest., 2008;118(5):1806-14.
    Species: Human
    Sample Types: Serum
  6. How a cytokine is chaperoned through the secretory pathway by complexing with its own receptor: lessons from interleukin-15 (IL-15)/IL-15 receptor alpha.
    Authors: Duitman EH, Orinska Z, Bulanova E, Paus R, Bulfone-Paus S
    Mol. Cell. Biol., 2008;28(15):4851-61.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Soluble Interleukin IL-15Ralpha is generated by alternative splicing or proteolytic cleavage and forms functional complexes with IL-15.
    Authors: Bulanova E, Budagian V, Duitman E, Orinska Z, Krause H, Ruckert R, Reiling N, Bulfone-Paus S
    J. Biol. Chem., 2007;282(18):13167-79.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Human T cells constitutively express IL-15 that promotes ex vivo T cell homeostatic proliferation through autocrine/juxtacrine loops.
    Authors: Miranda-Carus ME, Benito-Miguel M, Llamas MA, Balsa A, Martin-Mola E
    J. Immunol., 2005;175(6):3656-62.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. IL-15 and the initiation of cell contact-dependent synovial fibroblast-T lymphocyte cross-talk in rheumatoid arthritis: effect of methotrexate.
    Authors: Miranda-Carus ME, Balsa A, Benito-Miguel M, Perez de Ayala C, Martin-Mola E
    J. Immunol., 2004;173(2):1463-76.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Stimulation of T cell autoreactivity by anomalous expression of NKG2D and its MIC ligands in rheumatoid arthritis.
    Authors: Groh V, Bruhl A, El-Gabalawy H, Nelson JL, Spies T
    Proc. Natl. Acad. Sci. U.S.A., 2003;100(16):9452-7.
    Species: Human
    Sample Types: Serum

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